[One-stage repair of full-thickness skin defect at dorsal skin of middle phalanx fingers using artificial dermis combing with digital artery perforator fascial flaps].

OBJECTIVE: To explore clinical effects of the stageⅠrepair of full-thickness skin defect at dorsal skin of middle phalanx fingers using artificial dermis combing with digital artery perforator fascial flaps. METHODS: From January 2019 to May 2020, 21 patients(27 middle phalanx fingers)with full-thickness skin defect were repaired at stageⅠusing artificial dermis combing with digital artery perforator fascial flaps. All patients were emergency cases, and were accompanied by the exposure of bone tendon and the defects of periosteum and tendon membrane. Among patients, including 11 males and 10 females aged from 18 to 66 years old with an average age of (39.00±8.01) years old;9 index fingers, 10 middle fingers and 8 ring fingers;range of skin defect area ranged from (2.5 to 3.5) cm×(1.5 to 3.0) cm;range of exposed bone tendon area was (1.5 to 2.0) cm×(1.0 to 2.0) cm. The time from admission to hospital ranged from 1 to 6 h, operation time started from 3 to 8 h after injury. RESULTS: All patients were followed up from 6 to12 months with an average of (9.66±1.05) months. The wounds in 26 cases were completely healed at 4 to 6 weeks after operation. One finger has changed into wound infection with incompletely epithelialized dermis, and achieved wound healing at 8 weeks after dressing change. All fingers were plump with less scars. The healed wound surface was similar to the color and texture of the surrounding skin. These fingers have excellent wearability and flexibility. According to the upper limb function trial evaluation standard of Hand Surgery Society of Chinese Medical Association, the total score ranged from 72 to 100. 26 fingers got excellent result and 1 good. CONCLUSION: StageⅠrepair of full-thickness skin defect at dorsal skin of middle phalanx fingers using artificial dermis combing with digital artery perforator fascial flaps is easy to operate with less trauma. It has made satisfactory recovery of appearance and function of fingers. It could provide an effective surgical method for clinical treatment of full-thickness skin loss of fingers with tendon and bone exposure.

Fatty acid metabolism predicts prognosis and NK cell immunosurveillance of acute myeloid leukemia patients.

Background: Cell metabolic reprogramming is a hallmark of tumor prognosis, and fatty acid metabolism (FAM) plays a crucial role in the tumor microenvironment (TME). However, the relationship between FAM, TME, and prognosis of acute myeloid leukemia (AML) patients remains elusive. Methods: We extracted the single-cell RNA sequencing (scRNA-Seq) and bulk transcriptome data of AML patients from the TCGA and GEO databases and assessed the relationship between FAM, TME, and AML patient prognosis. We also performed functional enrichment (FE) assay to evaluate the significance of FAM in anti-AML immunosurveillance. Results: Our scRNA-Seq analysis revealed that the leukemic stem cell (LSC)-enriched population exhibited elevated levels of FAM-related genes. Using these FAM-related genes, we developed a prognostic model that accurately estimated AML patient outcome. FE analysis showed that FAM was strongly related to alterations of TME-based immunosurveillance in AML patients. More importantly, we demonstrated that FAM inhibition via pharmaceutical targeting of PLA2G4A, a highly expressed FAM gene in AML patients with poor prognosis, enhanced the NK cell-mediated immunosurveillance in leukemia cells. Conclusions: Leukemic stem cell (LSC)-enriched population exhibited elevated levels of FAM-related genes. We have successfully established the FAM formula that predicts AML patient prognosis and alterations in the TME-based immunosurveillance. We also found that PLA2G4A was a highly expressed FAM gene in AML patients with poor prognoses. Pharmaceutical targeting of PLA2G4A increased the expression of NKG2DL in leukemia cells in vitro and thus enhanced the NK cell-mediated immunosurveillance.

6-Phosphogluconolactonase Promotes Hepatocellular Carcinogenesis by Activating Pentose Phosphate Pathway.

Hepatocellular carcinoma (HCC) has a poor prognosis due to the rapid disease progression and early metastasis. The metabolism program determines the proliferation and metastasis of HCC; however, the metabolic approach to treat HCC remains uncovered. Here, by analyzing the liver cell single-cell sequencing data from HCC patients and healthy individuals, we found that 6-phosphogluconolactonase (PGLS), a cytosolic enzyme in the oxidative phase of the pentose phosphate pathway (PPP), expressing cells are associated with undifferentiated HCC subtypes. The Cancer Genome Atlas database showed that high PGLS expression was correlated with the poor prognosis in HCC patients. Knockdown or pharmaceutical inhibition of PGLS impaired the proliferation, migration, and invasion capacities of HCC cell lines, Hep3b and Huh7. Mechanistically, PGLS inhibition repressed the PPP, resulting in increased reactive oxygen species level that decreased proliferation and metastasis and increased apoptosis in HCC cells. Overall, our study showed that PGLS is a potential therapeutic target for HCC treatment through impacting the metabolic program in HCC cells.

[In vitro quantum dot-labeled rat bone marrow mesenchymal stem cells].

UNLABELLED: OBJECTIVE To explore the cytotoxicity, labeled time, marking rate, and effect on adhesion of quantum dot 655 (QD655) labeled rat bone marrow mesenchymal stem cells (BMSCs) in vitro, and to confirm its feasibility for stem cell labeling and tracer means for rat. METHODS: BMSCs were collected from the femur and tibia bone marrow cavity of a 2-week-old SD rat, cultured and identified. The 3rd passage of BMSCs were incubated with QD655 as the experimental group according to the recommended concentration of the markers. The cells were not labeled by QD655 as control group. The cell survival rate after QD655 labeling was detected by trypan-blue exclusion. The effect of QD655 on cell proliferation was observed by MTT. The osteogenic differentiation potential was identified by Alizarin red staining, alkaline phosphatase (ALP) staining, and real-time fluorogenic quantitative PCR. At immediately, 1, 2, 4, and 6 weeks, fluorescent microscopy was used to observe the labeled rate and scanning electron microscope was used to observe the cell adhesion to scaffold (bioglass/collagen composite). RESULTS: The cell survival rates were more than 90% in both experimental group and control group, showing no significant difference (P > 0.05). There was no significant difference in the cell proliferation between 2 groups (P > 0.05). Alizarin red staining and ALP staining showed positive results. Real-time fluorogenic quantitative PCR result showed that the mRNA expression levels of osteopontin, osteocalcin, collagen type I, ALP, and BMP-2 in the experimental group was significantly higher than those in the control group. The labeled rates were 96.50% +/- 1.59%, 93.30% +/- 1.51%, 72.40% +/- 2.90%, 40.10% + 3.60%, and 10.00% +/- 1.70% immediately, 1, 2, 4, and 6 weeks after labeling, respectively. The labeled rate in the control group was 0. Scanning electron microscope showed a good distribution of fusiform or polygonal cells in the pores of scaffold. CONCLUSION: QD655 can be used as a labeling marker for BMSCs. Rat BMSCs labeled with QD655 is of high efficiency and safety.

[Comparison of whole bone marrow culture method and density gradient centrifugation method of isolating hBMSCs].

OBJECTIVE: To make a comparative study on the effects of whole bone marrow culture method and density gradient centrifugation method in isolating hBMSCs. METHODS: hBMSCs were obtained from healthy adult volunteers and isolated by whole bone marrow culture method and density gradient centrifugation method. Primary cell morphology was observed using inverted phase contrast microscope and the cells in the 2nd passage were stained with HE after being cultured for 7 days. And then, the generation time of the primary, 2nd and 3rd passage hBMSCs was compared between two methods and the surface markers were detected by flow cytometer. In addition, the ALP expression in osteoinductive hBMSCs were evaluated by ALP activity kit at 3, 6 and 9 days and ALP staining was used for osteoinductive hBMSCs with Kaplow method at 9 days. RESULTS: Primary cells isolated with whole bone marrow culture method showed aggregation growth while cells isolated with density gradient centrifugation method showed diffusion growth. HE staining showed no significant difference in the morphology of the 2nd passage cells between these two methods. The generation time of primary cells isolated by whole bone marrow culture method (15.36 +/- 1.67) days was significantly shorter than that of cells isolated by density gradient centrifugation method [(18.57 +/- 1.05) days] (P < 0.01), while the generation time of the 2nd and 3rd passage cells showed no statistically significant differences between these methods (P > 0.05). The consent of positive surface markers (CD29, CD44, CD71, CD105, CD166) and negative surface marker CD34 in the 2nd cells showed no significant difference between these two isolation methods (P > 0.05); however, negative markers CD14 and CD45 showed significant difference (P < 0.01). The ALP expression in osteoinductive cells showed no statistical significant (P > 0.05) at 3, 6 and 9 days; and the ALP staining positive cell ratio of whole bone marrow culture method was basically in accordance with that of density gradient centrifugation method at 9 days. CONCLUSION: hBMSCs could be isolated by whole bone marrow culture method, and the cell isolation effects of whole bone marrow culture method are equivalent with density gradient centrifugation method.